Leukocytes often undergo rapid changes in cell phenotype, for example, from a resting to an activated state, which places significant metabolic demands on the cell. These rapid changes in metabolic ...demand need to be tightly regulated to support immune cell effector functions during the initiation and downregulation of an immune response. Prospects for implementing cancer immunotherapy also rest on the idea of optimizing the metabolic profile of immune cell effectors. Here, we examine this issue by focusing on neutrophils and NK cells as cells of increasing interest in cancer immunology and tumor immunometabolism, because they can be targeted or, in the case of NK, used as effectors in immunotherapy. In addition, neutrophils and NK cells have been shown to functionally interact. In the case of neutrophils, we also extended our interest to polymorphonuclear MDSC (PMN-MDSCs), since the granulocytic subset of MDSCs share many phenotypes and are functionally similar to pro-tumor neutrophils. Finally, we reviewed relevant strategies to target tumor metabolism, focusing on neutrophils and NK cells.
Prostate cancer (PCa) accounts as the most common non-cutaneous disease affecting males, and as the first cancer, for incidence, in male. With the introduction of the concept of immunoscore, PCa has ...been classified as a cold tumor, thus driving the attention in the development of strategies aimed at blocking the infiltration/activation of immunosuppressive cells, while favoring the infiltration/activation of anti-tumor immune cells. Even if immunotherapy has revolutionized the approaches to cancer therapy, there is still a window failure, due to the immune cell plasticity within PCa, that can acquire pro-tumor features, subsequent to the tumor microenvironment (TME) capability to polarize them. This review discussed selected relevant soluble factors transforming growth factor-beta (TGFβ), interleukin-6 (IL-6), IL-10, IL-23 and cellular components of the innate immunity, as drivers of tumor progression, immunosuppression, and angiogenesis within the PCa-TME.
Tissue regeneration or healing both require efficient vascularization within a tissue-damaged area. Based on this concept, a remarkable number of strategies, aimed at developing new tools to support ...re-vascularization of damaged tissue have emerged. Among the strategies proposed, the use of pro-angiogenic soluble factors, as a cell-free tool, appears as a promising approach, able to overcome the issues concerning the direct use of cells for regenerative medicine therapy. Here, we compared the effectiveness of adipose mesenchymal stem cells (ASCs), use as cell suspension, ASC protein extract or ASC-conditioned-medium (i.e., soluble factors), combined with collagenic scaffold, in supporting in vivo angiogenesis. We also tested the capability of hypoxia in increasing the efficiency of ASC to promote angiogenesis, via soluble factors, both in vivo and in vitro. In vivo studies were performed using the Integra® Flowable Wound Matrix, and the Ultimatrix in sponge assay. Flow cytometry was used to characterize the scaffold- and sponge-infiltrating cells. Real-time PCR was used to evaluate the expression of pro-angiogenic factors by stimulating Human Umbilical-Vein Endothelial Cells with ASC-conditioned media, obtained in hypoxic and normoxic conditions. We found that, in vivo, ACS-conditioned media can support angiogenesis similar to ASCs and ASC protein extract. Also, we observed that hypoxia increases the pro-angiogenic activities of ASC-conditioned media, compared to normoxia, by generating a secretome enriched in pro-angiogenic soluble factors, with bFGF, Adiponectine, ENA78, GRO, GRO-a, and ICAM1-3, as most regulated factors. Finally, ASC-conditioned media, produced in hypoxic condition, induce the expression of pro-angiogenic molecules in HUVECs. Our results provide evidence that ASC-conditioned-medium can be proposed as a cell-free preparation able to support angiogenesis, thus providing a relevant tool to overcome the issues and restrictions associated with the use of cells.
Abstract
Introduction. Natural Killer (NK) cells are innate lymphoid cells involved in tumour recognition/elimination. NK cells are altered in their phenotype and functions in diverse tumors, ...including prostate cancer (PCa). We demonstrated that PCa circulating NK cells (TANKs) acquire the pro-angiogenic/decidual-like CD56brightCD9+CD49a+ phenotype, release IL-8 and MMP-9, functionally support endothelial cell activation and secrete monocyte-recruiting/M2-like macrophage-polarizing factors.
Materials and methods. Here, we characterized the phenotype of tumour infiltrating NKs (TINKs) and tumour-associated (TANKs) in PCa patients and evaluated the contribution of STAT3, as possible driver of NK cell polarization. PCa TINKs and TANKs were characterized by multicolour flow cytometry (FC) for decidual-like surface markers (CD9, CD49a) and degranulation capabilities (CD107a). STAT3 activation, was investigated in circulating PCa NK cells, by FC. Using a drug-repurposing approach with the antipsychotic agent Pimozide (a chemical inhibitor of STAT3), we modulated STAT3 activation, ex vivo, in PCa TANKs and monitored their secretome changes, by commercially available protein membrane arrays together with their capability to degranulate and produce perforin/GranzymeB.
Results and discussion. We observed that PCa TINKs acquire the same CD9+CD49a+ decidual-like NK cell phenotype, as found in PCa TANKs. We detected the presence of CD56brightCD9+CD49a+ decidual-like NK cell also in peripheral blood of subjects with benign prostatic hyperplasia (BPH), but in a lower frequency, compared to those from PCa TANKs. Sera from PCa patients were enriched in IL-4, IL-6, IL-8 and IL-10, all cytokines able to activate STAT3 signalling. We detected increased phosphorylation of STAT3 in PCa TANKs, compared to NK cells from healthy controls, that was reduced following 24 hours of stimulation by Pimozide. This treatment resulted in decreased capabilities of PCa TANKs to secrete pro-angiogenic factors (IL-8, IL-6), molecules involved in monocytes recruitment/M2-like macrophage polarization (CCL-2, CCL5, GM-CSF, IL-10), together with increased degranulation, augmented secretion of (IFN-γ and TNF-α) and increased production of Perforin and Granzyme.
Conclusions. Our results suggest that STAT3 inhibition can be envisaged as a potential strategy to limit the generation of pro-angiogenic/decidual-like NKs, while contributing to NK cell re-education in PCa.
Citation Format: Matteo Gallazzi, Maria Teresa Palano, Martina Cucchiara, Federico Dehò, Paolo Capogrosso, Francesca Franzi, Fausto Sessa, Angelo Naselli, Lorenzo Mortara, Antonino Bruno. STAT-3 chemical inhibition modulates decidual-like polarization in NK cells from PCa patients and restore their anti-tumor activities. abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4595.
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